Using flow to improve clinical relevance in hepatocyte cultures
With Quasi Vivo®, Kirkstall’s flow system, you can quickly and easily set up and run in vitro assays to detect hepatotoxicity of novel compounds.
ASSESSMENT OF HEPATOXICITY USING QUASI VIVO®
- Drug induced liver injury (DILI) is a major cause of drug failure, both during clinical trials and after introduction to the market. A recent analysis of AstraZeneca project closures due to safety issues shows that 14% of preclinical failures and 12% of clinical failures were due to problems caused in the liver 1.
- Existing in vitro tests do not accurately mimic the physiological environment of the in vivo liver. One important missing factor is the presence of interstitial flow.
- Kirkstall’s Quasi Vivo® system allows the application of physiologically relevant flow rates. Evidence shows that key detoxification genes are up-regulated under flow conditions 2, liver organoids display typical liver morphology 3 and respond as expected to viability-decreasing compounds 4.
Many studies have shown that flow is important for correct physiological relevance of in vitro liver systems. Quasi Vivo® gives researchers the confidence that their experiments will translate into humans.
- Quasi Vivo® 900 try and accessories: tubing, reservoir bottles
- Parker 6 head peristaltic pump
- Inverted confical microscope for analysis
GENE EXPRESSION DATA
Changes in gene expression in human primary hepatocytes cultured under flow or static, compared to freshly isolated hepatocytes. Improvements in gene expression over static conditions are seen in Phase I and Phase II detoxification genes, as well as a transporter and xenobiotic transcription factor. Data taken from Vinci et al. 2.
Dose response curve for rat primary hepatocytes exposed to diclofenac under flow and static conditions. Flow enhances the response or sensitivity of the cells, and brings the culture environment closer to in vivo and correctly identifies diclofenac as a risk drug.
ACTIVITY OF KEY ENZYMES IN THE LIVER
Effect of medium flow on CYP-mediated monooxygenase activities. Shown are results from two independent cultures, in nmol h-1 106 cells-1. Although activities are lower than those observed in freshly isolated hepatocytes (FIH), Tolbutamide 4-hydroxylation, Midazolam 1-hydroxylation and Midazolam O-Glucuro were induced in two independent cultures.
Kirkstall’s Quasi Vivo® system provides a complete solution for the improvement of hepatocyte culture. Compared with traditional static culture techniques, genes are up-regulated and the response of cells to drugs is improved.
Along with our flow systems and pumps, we also provide a complete training and support package to help you through your experiments, from planning to execution. Quasi Vivo® is compatible with many 3D cell culture protocols, and can be integrated into your laboratory with ease.
This advanced in vitro technique is physiologically relevant, making your data more applicable to the human situation.
1. Cook, D. et al. Lessons learned from the fate of AstraZeneca’s drug pipeline: a five-dimensional framework. Nat. Rev. Drug Discov. 13, 419–31 (2014).
2. Vinci, B. et al. Modular bioreactor for primary human hepatocyte culture: Medium flow stimulates expression and activity of detoxification genes. Biotechnol. J. 6, 554–564 (2011).
3. Ramachandran, S. D. et al. In vitro generation of functional liver organoid-like structures using adult human cells. PLoS One 10, 1–14 (2015).
4. Rashidi, H., Alhaque, S., Szkolnicka, D., Flint, O. & Hay, D. C. Fluid shear stress modulation of hepatocyte-like cell function. Arch. Toxicol. 3–7 (2016). doi:10.1007/s00204-016-1689-8